scholarly journals In vitro Protein Synthesis and Measurement of the Stability of Messenger RNA in the Blue-green Alga, Anabaena variabilis

Microbiology ◽  
2000 ◽  
Vol 81 (1) ◽  
pp. 47-58 ◽  
Author(s):  
C. K. Leach ◽  
N. G. Carr
Keyword(s):  
Amino Acids ◽  
Green Alga ◽  
Messenger Rna ◽  
Antibiotic Sensitivity ◽  
Anabaena Variabilis ◽  
Blue Green Alga ◽  
Peptide Formation ◽  
A Cell ◽  
The Stability

A cell-free preparation of the blue-green alga, Anabaena variabilis, that incorporates 14C-labelled amino acids into protein has been prepared and characterized. The activation of amino acids to amino acid transfer RNAs was characterized, and assembly of these into peptides has been described with respect to cofactor requirements and antibiotic sensitivity. The properties of these systems and the effect of antibiotics on them are similar to those of bacteria. Both natural and synthetic messenger RNA were effective in peptide formation. The kinetics of incorporation of [14C]uracil into RNA has been examined and the stability of labelled RNA from A. variabilis measured by radioactivity loss and by its role in directing peptide synthesis. The half-life of messenger RNA from this organism is approximately 12 min, and as such is comparable to that of bacteria when based upon the mean generation time of the respective organisms.

Production of amino acids by the N2-fixing blue-green alga Anabaena variabilis var. kashiensis

1985 ◽  
Vol 140 (4) ◽  
pp. 333-339
Author(s):  
A.A. El-Essawy ◽  
E.Y. El-Ayouty ◽  
Y.M. El-Ayouty
Keyword(s):  
Amino Acids ◽  
Green Alga ◽  
Anabaena Variabilis ◽  
Blue Green Alga

Effects of amino acids on the blue-green alga (cyanobacteria) Tolypothrix scytonemoides (Gardner) Geitler

2006 ◽  
Vol 8 (3) ◽  
pp. 286-293
Author(s):  
Uma Maheshwari Rajendran ◽  
Elango Kathirvel ◽  
Anand Narayanaswamy
Keyword(s):  
Amino Acids ◽  
Green Alga ◽  
Blue Green Alga

Using FRET to Measure the Time it Takes for a Cell to Destroy a Virus

2019 ◽  
Author(s):  
Candace E. Benjamin ◽  
Zhuo Chen ◽  
Olivia Brohlin ◽  
Hamilton Lee ◽  
Stefanie Boyd ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Rhodamine B ◽  
Virus Like Particle ◽  
A Cell ◽  
Viral Nanoparticles ◽  
The Stability ◽  
Open Question ◽  
Viral Surface ◽  
Fret Pair

<div><div><div><p>The emergence of viral nanotechnology over the preceding two decades has created a number of intellectually captivating possible translational applications; however, the in vitro fate of the viral nanoparticles in cells remains an open question. Herein, we investigate the stability and lifetime of virus-like particle (VLP) Qβ - a representative and popular VLP for several applications - following cellular uptake. By exploiting the available functional handles on the viral surface, we have orthogonally installed the known FRET pair, FITC and Rhodamine B, to gain insight of the particle’s behavior in vitro. Based on these data, we believe VLPs undergo aggregation in addition to the anticipated proteolysis within a few hours of cellular uptake.</p></div></div></div>

scholarly journals InsP7is a small-molecule regulator of NUDT3-mediated mRNA decapping and processing-body dynamics

2020 ◽  
Vol 117 (32) ◽  
pp. 19245-19253 ◽  
Author(s):  
Soumyadip Sahu ◽  
Zhenzhen Wang ◽  
Xinfu Jiao ◽  
Chunfang Gu ◽  
Nikolaus Jork ◽  
...  
Keyword(s):  
Stress Responses ◽  
Messenger Rna ◽  
Control Process ◽  
Stem Cell Differentiation ◽  
Wild Type ◽  
Inositol Pyrophosphate ◽  
Body Dynamics ◽  
Processing Body ◽  
The Stability

Regulation of enzymatic 5′ decapping of messenger RNA (mRNA), which normally commits transcripts to their destruction, has the capacity to dynamically reshape the transcriptome. For example, protection from 5′ decapping promotes accumulation of mRNAs into processing (P) bodies—membraneless, biomolecular condensates. Such compartmentalization of mRNAs temporarily removes them from the translatable pool; these repressed transcripts are stabilized and stored until P-body dissolution permits transcript reentry into the cytosol. Here, we describe regulation of mRNA stability and P-body dynamics by the inositol pyrophosphate signaling molecule 5-InsP7(5-diphosphoinositol pentakisphosphate). First, we demonstrate 5-InsP7inhibits decapping by recombinant NUDT3 (Nudix [nucleoside diphosphate linked moiety X]-type hydrolase 3) in vitro. Next, in intact HEK293 and HCT116 cells, we monitored the stability of a cadre of NUDT3 mRNA substrates following CRISPR-Cas9 knockout ofPPIP5Ks(diphosphoinositol pentakisphosphate 5-kinases type 1 and 2, i.e.,PPIP5KKO), which elevates cellular 5-InsP7levels by two- to threefold (i.e., within the physiological rheostatic range). ThePPIP5KKO cells exhibited elevated levels of NUDT3 mRNA substrates and increased P-body abundance. Pharmacological and genetic attenuation of 5-InsP7synthesis in the KO background reverted both NUDT3 mRNA substrate levels and P-body counts to those of wild-type cells. Furthermore, liposomal delivery of a metabolically resistant 5-InsP7analog into wild-type cells elevated levels of NUDT3 mRNA substrates and raised P-body abundance. In the context that cellular 5-InsP7levels normally fluctuate in response to changes in the bioenergetic environment, regulation of mRNA structure by this inositol pyrophosphate represents an epitranscriptomic control process. The associated impact on P-body dynamics has relevance to regulation of stem cell differentiation, stress responses, and, potentially, amelioration of neurodegenerative diseases and aging.

scholarly journals Chemical aminoacylation of tRNAs with fluorinated amino acids for in vitro protein mutagenesis

2010 ◽  
Vol 6 ◽  
Author(s):  
Shijie Ye ◽  
Allison Ann Berger ◽  
Dominique Petzold ◽  
Oliver Reimann ◽  
Benjamin Matt ◽  
...  
Keyword(s):  
Amino Acids ◽  
Translation System ◽  
Biologically Relevant ◽  
Fluorinated Amino Acids ◽  
Free Translation ◽  
Trna Species ◽  
A Cell ◽  
Protein Environment ◽  
Vitro Protein

This article describes the chemical aminoacylation of the yeast phenylalanine suppressor tRNA with a series of amino acids bearing fluorinated side chains via the hybrid dinucleotide pdCpA and ligation to the corresponding truncated tRNA species. Aminoacyl-tRNAs can be used to synthesize biologically relevant proteins which contain fluorinated amino acids at specific sites by means of a cell-free translation system. Such engineered proteins are expected to contribute to our understanding of discrete fluorines’ interaction with canonical amino acids in a native protein environment and to enable the design of fluorinated proteins with arbitrary desired properties.

scholarly journals Comparison of four methods of differential typing of isolates ofShigella sonnei

1981 ◽  
Vol 87 (3) ◽  
pp. 339-355 ◽  
Author(s):  
S. Helgason ◽  
D. C. Old
Keyword(s):  
Antibiotic Sensitivity ◽  
The Other ◽  
Drug Resistant ◽  
Typing Method ◽  
Stable Property ◽  
Discriminating Power ◽  
Sensitivity Tests ◽  
The Stability

SummaryAn epidemiological study of Sonne dysentery in Dundee during the years 1971–6 was made by examining, in respect of 1420 isolates ofShigella sonnei, the discriminating power of colicine typing, antibiogram testing, biotyping and resistotyping and the stability of the markers they provided.Colicine typing identified nine colicine types, including four not previously described. However, because types 4 and 4 var., determined bycolIb, and type U, producing no colicines, accounted for 96 % of the isolates, discrimination with colicine typing was poor. In antibiotic sensitivity tests, 13 different antibiogram patterns were noted. Less than 1 % of the isolates were sensitive to all of the eight antibiotics tested; most were multiply drug-resistant. Resistance to kanamycin, neomycin and paromomycin (KNP) was apparently due to a single resistance determinant, widely distributed in a majority (53%) of the isolates. When definitive times were chosen for reading each biotyping test, only maltose and rhamnose of the 13 ‘sugars’ tested differentiated isolates into prompt- and late-fermenting types. Though the ability to ferment rhamnose was a stable property, it discriminated only 1·5% of the minority, late-fermenting type. Resistotyping with six chemicals discriminated eight epidemiologically valid resistotypes, including three new types. However, 93 % of the isolates belonged to only three resistotypes.Analysis of the data for isolates from 286 epidemiologically distinct episodes showed that the variability of colicine and antibiogram characters, found among isolates within, respectively, 40 and 28 % of the episodes, was generally associated with loss or gain of a plasmid (‘colIb-KNP’) which determined production of colicine Ib and KNP resistance. These characters varied bothin vivoandin vitro. Variability of resistotype characters, on the other hand, was observed in only 28 (9%) episodes, 14 of which possibly represented examples of mixed or sequential infections.For accurate epidemiological tracing of strains ofSh. sonneiin a community, resistotyping, the technique showing the greatest discrimination and least variability of the four tested, should be included as the principal typing method.

scholarly journals PTP1B Modulates the Association of β-Catenin with N-cadherin through Binding to an Adjacent and Partially Overlapping Target Site

2002 ◽  
Vol 277 (51) ◽  
pp. 49989-49997 ◽  
Author(s):  
Gang Xu ◽  
Carlos Arregui ◽  
Jack Lilien ◽  
Janne Balsamo
Keyword(s):  
Amino Acids ◽  
Cell Surface ◽  
Tyrosine Phosphatase ◽  
Target Site ◽  
Target Domain ◽  
Chick Retina ◽  
Cell Permeable Peptide ◽  
A Cell ◽  
Relationship Of

The nonreceptor tyrosine phosphatase PTP1B associates with the cytoplasmic domain of N-cadherin and may regulate cadherin function through dephosphorylation of β-catenin. We have now identified the domain on N-cadherin to which PTP1B binds and characterized the effect of perturbing this domain on cadherin function. Deletion constructs lacking amino acids 872–891 fail to bind PTP1B. This domain partially overlaps with the β-catenin binding domain. To further define the relationship of these two sites, we used peptides to competein vitrobinding. A peptide representing the most NH2-terminal 8 amino acids of the PTP1B binding site, the region of overlap with the β-catenin target, effectively competes for binding of β-catenin but is much less effective in competing PTP1B, whereas two peptides representing the remaining 12 amino acids have no effect on β-catenin binding but effectively compete for PTP1B binding. Introduction into embryonic chick retina cells of a cell-permeable peptide mimicking the 8 most COOH-terminal amino acids in the PTP1B target domain, the region most distant from the β-catenin target site, prevents binding of PTP1B, increases the pool of free, tyrosine-phosphorylated β-catenin, and results in loss of N-cadherin function. N-cadherin lacking this same region of the PTP1B target site does not associate with PTP1B or β-catenin and is not efficiently expressed at the cell surface of transfected L cells. Thus, interaction of PTP1B with N-cadherin is essential for its association with β-catenin, stable expression at the cell surface, and consequently, cadherin function.

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